• Journal of Microbiology and Biotechnology
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• v.4 no.2
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• pp.119-125
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• 1994

Steroid $\Delta^1$-dehydrogenase purified from hydrocortisone-induced cells of Arthrobacter simplex converted various 3-ketosteroids into their corresponding $\Delta^1$-dehydrogenated products. The transformation efficiencies depend upon the chemical structure of the steroids, especially length of the side chain at 17 position and hydroxyl groups at 11 and 17 positions. The Km values for androstenedione, the most favorable substrate examined, and hydrocortisone were 74 ${\mu}M$ and 294 ${\mu}M$, respectively. The optimum temperature and pH of the enzyme reaction were 35$^{\circ}C$ and pH 9, respectively, and the enzyme was relatively stable at the range from 20 to 35$^{\circ}C$ and from pH 5 to 10 after one hour of incubation. The enzyme activity was markedly inhibited in the presence of $Cu^{2+},\;Fe^{3+},\;Hg^{2+},\;Mo^{6+}$ ions, and somewhat inhibited by $Zn^{2+}$ and $Fe^{2+}$. $\alpha,\alpha'$-Dipyridyl that inhibits 9$\alpha$-hydroxylase and accumulates 1,4-androstadiene-3,17-dione from sterols revealed no inhibitory effect on this enzyme. EGTA showed inhibitory effect. $\beta$-Estradiol competitively inhibited the enzyme activity. Chemical modifications of the enzyme were attempted with several reagents. p-Hydroxymer-curibenzoate showed inhibition of the enzyme activity and protection of the substrate. This suggests that cysteine residue may be involved in the active site of the enzyme.

• Journal of Physiology & Pathology in Korean Medicine
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• v.25 no.3
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• pp.417-424
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• 2011

Ketoconazole (KET) is an antifungal drug with a broad spectrum of activity that also induces reproductive toxicity in humans and animals. KET inhibits C17-20 lyase which blocks the conversion of 17 ${\alpha}$-hydroxyprogesterone to androstenedione. The effect of Cuscutae semen(CS) extract against KET-induced testicular damage was evaluated in male rats. CS extract was administered orally (100 and 200 mg/kg) for 26 days. Three weeks after CS extract administration, KET was CS-administered intraperitoneally at a dose of 100 mg/kg once a day for 5 days. KET-induced reproductive toxicity was associated with clear reductions of the weights of testes and epididymides, sperm indices and serum testosterone levels. In addition, marked oxidative damage to testicular lipids and alterations of natural antioxidant enzymes were reported in association with KET toxicity. Most of the KET-induced effects were greatly decreased with the concomitant application of CS extract. This study suggests a protective role of Cuscutae semen extract that could be attributed to its antioxidant properties.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.47 no.6
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• pp.882-887
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• 2014

We studied oocyte steroidogenesis in blacktip grouper Epinephelus fasciatus ovarian follicles during vitellogenesis. Vitellogenic oocytes with average diameters of 0.45, 0.48 and 0.50 mm were incubated in vitro in the presence of $[^3H]17{\alpha}$-hydroxyprogesterone as a precursor. The steroid metabolites were analyzed using thin layer chromatography (TLC), high performance liquid chromatography (HPLC), and gas chromatography-mass spectrometry (GC/MS). The major metabolites in the vitellogenic oocytes were androstenedione ($A_4$), testosterone (T), estradiol-$17{\beta}$ ($E_2$), and estrone ($E_1$). The metabolites of androgen ($A_4$ and T) were higher in the 0.50-mm oocytes than in the 0.45- and 0.48-mm oocytes, while the estrogen metabolites (E2 and E1) were lower in the 0.50-mm oocytes. These results suggest that 0.50-mm oocytes are fully vitellogenic following initiation of the maturation process.

• Journal of Ginseng Research
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• v.9 no.2
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• pp.213-220
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• 1985

It was attempted to observe the effects of ginseng saponin, one of the major components of the roots of Panax ginseng, on androgen biosynthesis from cholesterol in vitro as well as in vivo in rat testis. Ginseng saponin was administered by stomach tubing prior to intraperitoneal injection of cholesterol containing (4-14C)-cholesteroll into adult male rats and the liver, testis and blood serum were analyzed. The first high radioactivity of the liver and blood serum of test animal was observed at 6 hours after radioactive cholesterol injection, while that of control appeared at 12 hours after the injection. In the case of testis, the first high radioactivity of test group appeared between 4 and 6 hours after the radioactive cholesterol injection, while that of control appeared at 10-14 hours. Analysis of radioactivity distribution of cholesterol, androstenedione and testosterone in the testis of rats fed with/without ginseng saponin piror to (4-14C)-cholesterol injection showed that the saponin stmulated the synthesis of androgens from cholesterol. This was confirmed again by in vitro experiment using testis homogenate as an enzyme source. From the above experimental results, it was suggested that the ginseng saponin stimulates both cholesterol transport and the biosynthesis of androgens from cholesterol in rat testis.

• The Journal of Korean Obstetrics and Gynecology
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• v.27 no.4
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• pp.25-42
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• 2014

Objectives: To observe the effects of Herb medication on Polycystic Ovary (PCO) in rats. Methods: Riss (Research Information Sharing Service) was searched for Oriental medicine about PCO on 2000~2014. So, Experimental studies using PCO model induced by Estradiol Valerate (EV) was selected to analyze how effects they were. Consequently, 10 studies published on 2000~2014 were selected to analyze about weight of model and both ovaries, the number aries, the number of follicle and corpus of follicle and corpus luteum, the level of serum Androstenedione (ADD) and total estrogen, the level of Nerve Growth Factor (NGF) and Corticotropin Releasing Factor (CRF), the crossover rate and the number of implantation. Results: Among them, there was significantly difference between treatment group and control group in weight of both ovaries, the number of follicle and corpora lutea, the crossover rate and the number of implantation. Conclusions: The effects of Herb medication was observed on making weight of ovaries with PCO to promote. The study about effect of Herb medication on NGF and CRF with PCOS had to be conducted. The further research have to focus on treatment of subfertility and infertility, that is field showing the advantage of Oriental medicine.

• Development and Reproduction
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• v.25 no.2
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• pp.75-82
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• 2021

We studied steroid metabolites produced from red-spotted grouper ovarian follicles during maturation. Oocytes with 350-500 ㎛ diameter were in vitro incubated in the presence of [³H] 17α-hydroxyprogesterone as a precursor. Steroid metabolites were extracted from incubated media and oocytes. The extracts were separated and identified using thin layer chromatography, high performance liquid chromatography and gas chromatography-mass spectrometry. The identified metabolites were androstenedione (A₄), testosterone (T) and estrone (E₁). The metabolites of A₄ was dominant in all size of oocytes and it was the highest in 480 ㎛ diameter oocytes. The metabolites of two progestins, 17α,20β-dihydroxy-4-pregnen-3-one and 17α,20α-dihydroxy-4-pregnen-3-one were detected in the oocytes less than 480 ㎛ diameter although they were not identified definitely. In the oocytes of 480 ㎛ diameter, metabolite of progestin was the highest, and germinal vesicle (GV) was still in the middle of cytoplasm. In the oocytes of 500 ㎛ diameter, GV was began to migrate and the major metabolites were A₄ and E₁. The metabolite of E₁ was detected in all size of oocytes and it was higher than that of E₂. These results suggest that oocytes of 480 ㎛ diameter are the transitional stage involving steroidogenic shift to final oocyte maturation and potential function of E₁ during maturation process.

• Development and Reproduction
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• v.17 no.4
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• pp.421-426
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• 2013

In this study, oocyte steroidogenesis are investigated in relation to oocyte development in the starry flounder, Platichthys stellatus, a marine multiple spawner. Vitellogenic (0.52 and 0.55 mm oocyte diameter) and mature oocytes (0.63, 0.66 and 0.71 mm oocyte diameter) were incubated in vitro in the presence of $[^3H]17{\alpha}$-hydroxyprogesterone ($[^3H]17{\alpha}$-OHP) as a precursor. Steroid metabolites were extracted from the incubated media and oocytes, the extracts were separated and identified by thin-layer chromatography (TLC), high performance liquid chromatography (HPLC) and gas chromatographymass spectrometry (GC-MS). The major metabolites produced from $[^3H]17{\alpha}$-OHP were androgens [androstenedione ($A_4$) and testosterone (T)] and estrogens [$17{\beta}$-estradiol ($E_2$) and estrone ($E_1$)] and progestins [$17{\alpha},20{\alpha}$-dihydroxy-4-pregnen-3-one ($17{\alpha}20{\alpha}P$) and $17{\alpha},20{\beta}$-dihydroxy-4-pregnen-3-one ($17{\alpha}20{\beta}P$)] in vitellogenic and mature oocytes. The results from this study suggest the potential roles of $E_1$ in the oocytes with diameter 0.52-0.71 mm, $17{\alpha}20{\alpha}P$ and $17{\alpha}20{\beta}P$ at the oocytes of 0.63, 0.66 and 0.71 mm.

• Journal of Acupuncture Research
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• v.23 no.3
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• pp.1-19
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• 2006

Objectives : This study was carried out to determine the effects of Gamidokwalgisaeng-tang(GDS) oral administration and electro-acupuncture at $BL_{52}$ & $GB_{39}$ on the postmenopausal osteoporosis. Methods: In order to study the effects of the Gamidokwalgisaeng-tang(GDS) oral administration and electro-acupuncture at $BL_{52}$, $GB_{39}$ on osteoporotic rats induced by ovariectomy, body weight, femur weight, uterine weight, serum level of osteocalcin, alkaline phosphatase(ALP), cholesterol, triglyceride, estradiol, androstenedione, testosterone and urinary excretions of calcium & phosphorus were measured. Results : 1. Body weight in all experimental group decreased significantly in comparison with ovariectomized group(OVX). 2. The weight of femur compared with body increased significantly in GDS oral administration and combination treatment group in comparison with ovariectomized group(OVX). 3. Uterine endometrial morphology is caused by ovariectomy and the weight of uterus decreased significantly in control and all experimental group in comparison with sham. 4. The serum level of osteocalcin decreased in all experimental group in comparison with OVX, but significantly in electro-acupuncture and combination treatment group. 5. The serum alkaline phosphatase(ALP) activity decreased in electro-acupuncture and combination treatment group in comparison with OVX, but only significantly in electro-acupuncture group. 6. The serum level of cholesterol increased significantly in OVX in comparison with sham and decreased in all experimental group in comparison with OVX, not insignificantly. The serum level of triglyceride increased in all experimental group in comparison with OVX, not insignificantly. 7. Urinary excretions of calcium and phosphorus decreased significantly in all experimental group in comparison with OVX. Conclusion : We suggest that GDS oral administration and electro-acupuncture at $BL_{52}$ & $GB_{39}$ are available for prevention and curing about the postmenopausal osteoporosis.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.48 no.4
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• pp.483-488
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• 2015

We studied oocyte steroidogenesis in the blackfin flounder Glyptocephalus stelleri as a region-specific species, in the East Sea of Korea during the spawning season. Maturing oocytes (0.76, 0.82, 0.88, and 0.91 mm in oocyte diameter) were incubated in vitro in the presence of [$^3H$] $17{\alpha}$-hydroxyprogesterone ($[^3H]17{\alpha}$-OHP) as a precursor. Steroid metabolites were extracted from the incubated medium and oocytes, and the extracts were separated and identified by thin-layer chromatography (TLC), high-performance liquid chromatography (HPLC) and gas chromatographymass spectrometry (GC/MS). The major metabolites produced from $[^3H]17{\alpha}$-OHP were androgens [androstenedione (A4) and testosterone (T)] and estrogens [$17{\beta}$-estradiol (E2) and estrone (E1)] and progestins [$17{\alpha},20{\alpha}$-dihydroxy-4-pregen-3-one ($17{\alpha}20{\alpha}P$) and $17{\alpha}20{\beta}$-dihydroxy-4-pregnen-3-one ($17{\alpha}20{\beta}P$)] in maturing oocytes. The metabolic rate of $17{\alpha}20{\beta}$ was elevated (29.04%) in oocytes measuring 0.88 mm (nucleus migration stage following the induction of germinal vesicle breakdown), but was very low in oocytes measuring 0.76, 0.82, and 0.91 mm (0.42, 0.67, and 2.62%, respectively). From these results, we suggest that $17{\alpha}20{\beta}P$ acts as a maturation-inducing steroid in the blackfin flounder.

• Asian-Australasian Journal of Animal Sciences
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• v.12 no.3
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• pp.348-353
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• 1999

Ten Australian Stock Horses colts (five yearling and five 3-year old colts) of which 2 yearlings and 2 three year old colts served as control animals while 3 yearlings and 3 three year old colts received two GnRH immunizations within 4 weeks interval were used in this study. By the 5th to 6th week after immunization, the GnRH antibody titres in the plasma rose above 1:1000 and attained peak levels of 1:6500 by the 8th week and gradually declined to about 1:3000 by the 10th week in both the age groups. The testosterone and androstenedione concentrations of the control colts in both age groups were significantly greater (p<0.05) than that of the vaccinated groups. During the immunosuppression period, the vaccinated colts behaved like geldings. Semen could not be collected from 2 of the 3 three-year old vaccinated colts. The testicular dimensions, testicular weight, parenchymal weight, seminiferous tubule volumes, interstitial space volumes, Leydig cell volume, seminiferous tubule % of the control colts were significantly greater than those of the vaccinated colts in both the age groups. The 3-year old control colts had a significantly (p<0.05) greater % of Leydig cells than the control and vaccinated 1-year old colts. There was arrest of spermatogenesis with complete absence of sperm in the testes of the vaccinated colts while there was various stages of spermatogenesis in those of the control colts. Morphometric analysis demonstrated that the 3-year old colts had significantly (p<0.05) greater DSP/gm of testis and DSP/testis than those of the 1-year old control colts. This study elucidated that the GnRH immunization could suppress the testicular function of the 3-year old and yearling colts.